Methods to treat conditions mediated by chemokine receptors

ABSTRACT

The present invention is drawn to novel antiviral compounds, pharmaceutical compositions and their use. More specifically this invention is drawn to derivatives of monocyclic polyamines which have activity in standard tests against HIV-infected cells as well as other biological activity related to binding of ligands to chemokine receptors that mediate a number of mammalian embryonic developmental processes.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. Ser. No. 10/143,692filed 9 May 2002, which is a continuation of U.S. Ser. No. 09/111,895filed 8 Jul. 1998, now U.S. Pat. No. 6,506,770.

TECHNICAL FIELD

[0002] The present invention is drawn to novel antiviral compounds,pharmaceutical compositions and their use. More specifically thisinvention is drawn to derivatives of monocyclic polyamines which haveactivity in standard tests against HIV-infected cells as well as otherbiological activity related to binding of ligands to chemokine receptorsthat mediate a number of mammalian embryonic developmental processes.

BACKGROUND ART

[0003] Approximately 40 human chemokines have been described, thatfunction, at least in part, by modulating a complex and overlapping setof biological activities important for the movement of lymphoid cellsand extravasation and tissue infiltration of leukocytes in response toinciting agents (see, for example: Ponath, P., Exp. Opin. Invest. Drugs(1998) 7:1-18). These chemotactic cytokines, or chemokines, constitute afamily of proteins, approximately 8-10 kDa in size. Chemokines appear toshare a common structural motif, that consists of 4 conserved cysteinesinvolved in maintaining tertiary structure. There are two majorsubfamilies of chemokines: the “CC” or β-chemokines and the “CXC” orα-chemokines. The receptors of these chemokines are classified basedupon the chemokine that constitutes the receptor's natural ligand.Receptors of the β-chemokines are designated “CCR”; while those of theα-chemokines are designated “CXCR”.

[0004] Chemokines are considered to be principal mediators in theinitiation and maintenance of inflammation. More specifically,chemokines have been found to play an important role in the regulationof endothelial cell function, including proliferation, migration anddifferentiation during angiogenesis and re-endothelialization afterinjury (Gupta, et al., J. Biolog. Chem. (1998) 7:4282-4287). Twospecific chemokines have been implicated in the etiology of infection byhuman immunodeficiency virus (HIV).

[0005] In most instances, HIV initially binds via its gp120 envelopeprotein to the CD4 receptor of the target cell. A conformational changeappears to take place in the gp120 which results in its subsequentbinding to a chemokine receptor, such as CCR-5 (Wyatt, et al., Science(1998) 280:1884-1888). HIV-1 isolates arising subsequently in theinfection bind to the CXCR-4 chemokine receptor. In view of the factthat the feline immunodeficiency virus, another related retrovirus,binds to a chemokine receptor without needing to bind first to the CD4receptor, suggests that chemokine receptors may be the primordialobligate receptors for immunodeficiency retroviruses.

[0006] Following the initial binding by HIV to CD4, virus-cell fusionresults, which is mediated by members of the chemokine receptor family,with different members serving as fusion cofactors for macrophage-tropic(M-tropic) and T cell line-tropic (T-tropic) isolates of HIV-1 (Carroll,et al., Science (1997) 276: 273-276). During the course of infectionwithin a patient, it appears that a majority of HIV particles shift fromthe M-tropic to the more aggressive T-tropic viral phenotype (Miedema,et al., Immune. Rev. (1994) 140:35). Curiously, the M-tropic viralphenotype correlates with the virus's ability to enter the cellfollowing binding of the CCR-5 receptor, while the T-tropic viralphenotype correlates with viral entry into the cell following bindingand membrane fusion with the CXCR-4 receptor. Clinically observationssuggest that patients who possess genetic mutations in the CCR-5 orCXCR-4 appear resistant or less susceptible to HIV infection.

[0007] However, the binding of chemokine receptors to their naturalligands appears to serve a more evolutionary and central role than onlyas mediators of HIV infection. The chemokine receptor, CXCR-4 has beenfound to be essential for the vascularization of the gastrointestinaltract (Tachibana, et al., Nature (1998) 393:591-594) as well ashematopoiesis and cerebellar development (Zou, et al., Nature (1998)393:591-594). Interference with any of these important functions servedby the binding of pre-B-cell growth-stimulating factor/stromal derivedfactor (PBSF/SDF-1) to the CXCR-4 chemokine receptor results in lethaldeficiencies in vascular development, hematopoiesis and cardiogenesis.Similarly, fetal cerebellar development appears to rely upon theeffective functioning of CXCR-4 in neuronal cell migration andpatterning in the central nervous system. This G-protein-coupledchemokine receptor appears to play a critical role in ensuring thenecessary patterns of migration of granule cells in the cerebellaranlage.

[0008] In attempting to better understand the relationship betweenchemokines and their receptors, recent experiments to block the bindingof HIV to the CXCR4 chemokine receptor were carried out through the useof monoclonal antibodies or small molecules that appear to suggest auseful therapeutic strategy (Schols, et al., J. Exp. Med. (1997)186:1383-1388; Schols, et al., Antiviral Research (1997) 35:147-156).Small molecules, such as bicyclams, appear to specifically interferewith the CXCR-4 binding and not CCR-5 binding (Donzella, et al., NatureMedicine (1998) 4:72-77). These experiments demonstrated interferencewith HIV entry and membrane fusion into the target cell in vitro.Additional experiments monitoring the calcium flux or Ca²⁺ mobilizationassay demonstrated that a bicyclam also functioned as an antagonist tosignal transduction resulting from the binding of stromal derived factoror SDF-1α, the natural chemokine to CXCR-4.

[0009] U.S. Pat. No. 5,583,131, U.S. Pat. No. 5,698,546 and allowedcopending U.S. application Ser. No. 08/659,500 disclose cyclic compoundsthat are active against HIV-1 and HIV-2 in in vitro tests. We have nowdiscovered that these compounds exhibit anti-HIV activity due to theirbinding to the chemokine receptor 4 (CXCR-4 or Fusin receptor),expressed on the surface of certain cells of the immune system. Thiscompetitive binding thereby protects these target cells from infectionby HIV which utilize the CXCR-4 receptor for entry. We have discoveredthat the disclosed compounds also antagonize the binding, signaling andchemotactic effects of the natural CXC-chemokine for CXCR-4, stromalcell-derived factor 1α (SDF-1α). Herein, we further disclose novelcompounds that demonstrate protective effects against HIV infection oftarget cells by inhibition of binding in vitro to the CC-5 receptor(CCR-5).

DISCLOSURE OF THE INVENTION

[0010] The present invention provides novel compounds, that demonstrateprotective effects on target cells from HIV infection as well asdemonstrate other biological activities related to the ability of thesecompounds to inhibit the binding by the natural ligand to its chemokinereceptor.

[0011] Accordingly, the present invention provides a macrocycliccompound of formula I:

V-CR¹R²—Ar—CR³R⁴—N(R⁵)—(CR⁶R⁷)_(x)—R⁸  (I)

[0012] wherein V is a cyclic polyamine moiety having a total of 9 to 24members and from 3 to 6 optionally substituted amine nitrogens spaced bytwo or more optionally substituted carbon atoms from each other, andwhich may optionally comprise a fused aromatic or heteroaromatic ring;

[0013] R¹ to R⁷ may be the same or different and are independentlyselected from hydrogen or straight, branched or cyclic C₁₋₆ alkyl;

[0014] R⁸ is a heterocyclic group, a substituted aromatic group, or amercaptan group;

[0015] Ar is an aromatic or heteroaromatic ring each optionallysubstituted at single or multiple positions with electrons-donating orwithdrawing groups;

[0016] x is 1 or 2;

[0017] and the acid addition salts and metal complexes thereof.

[0018] Preferably V is a 14- to 17-membered fused or unfused ringsystem, such as a cyclam system or a4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene system ora derivative thereof, and especially a cyclam system or derivativethereof. The moiety V may be substituted at C or N non-linking atoms,suitably by hydroxyl, alkoxy, thiol, thioalkyl or any other atom orgroup which does not adversely affect the activity or toxicity of thecompounds but may reduce the basicity of the amines, for examplehalogen, nitro, carboxy, carboxyamido, sulphonic acid or phosphate.Suitably the fused aromatic or heteroaromatic ring is phenyl, pyridine,pyrimidine, pyrazine, imidazole or thiazole. Preferably, the fusedaromatic or heteroaromatic ring is phenyl or pyridine.

[0019] Preferably R¹ to R⁷ are each hydrogen.

[0020] Preferably R⁸ is selected from pyridine, pyrimidine, pyrazine,imidazole, thiophene, thiophenyl, aminobenzyl, piperidinyl, piperazinylor a mercaptan group.

[0021] Preferably Ar is phenyl. Preferred substituents are alkyl, aryl,amino, alkoxy, hydroxy, halogen, carboxyl and carboxamido.

[0022] The invention also includes what may be termed as “pro-drug”,that is protected forms of the compounds, which release the compoundafter administration to a patient. For example, the compound may carry aprotective groups which is split off by hydrolysis in body fluids e.g.in the bloodstream, thus releasing active compound or are oxidized orreduced in body fluids to release the compound. A discussion orpro-drugs may be found in Smith and Williams' Introduction to thePrinciples of Drug Design, H. J. Smith, Wright, Second Edition, London1988.

[0023] Acid addition salts, for example hydrochlorides, and non-toxiclabile metal complexes of compounds of formula I are also activecompounds according to the present invention. Non-toxic in the presenttense has to be considered with reference to the prognosis for theinfected patient without treatment. Copper and zinc complexes arepreferred although other metals such as nickel may be considered,whereas less labile metals such as cobalt and rhodium are less preferredbecause of likely lower selectivity.

[0024] Compounds of formula (I) are novel. Accordingly, a further aspectof the invention provides a process for the preparation of a compound offormula (I) which comprises the following steps:

[0025] (i) nucleophilic attack by the cyclic polyamine V having a singleunprotected amine nitrogen, all other amine nitrogen atoms beingprotected, on an excess of a compound of formula (II)

Y—CR¹R²—Ar—CR³R⁴—Y  (II)

[0026] wherein R¹ to R⁴ and Ar are as hereinbefore defined, and each Yis an active substituent which can be displaced by the unprotectednitrogen of polyamine V and is preferably selected from Br, Cl, I,methane sulphonate, 4-toluenesulphonate, trifluoromethane sulphonate.

[0027] It is well within the capabilities and knowledge of the skilledsynthetic chemist to protect the amine nitrogens of cyclic polyamines,and it is preferred to use substitution by methanesulphonyl and/ortoluenesulphonyl and/or diethoxyphosphoryl (see: Bridger, et al., J.Med. Chem. (1995) 38:366-378; Bridger, et al., U.S. Pat. No. 5,583,131or Bridger et al., U.S. Pat. No. 5,698,546) and/or nitrobenzenesulfonyl(Fukuyama, et at., Tetrahedron Letters (1995) 36:6373-6374.

[0028] The protected polyamine V is firstly reacted with a 5- to 10-foldexcess of a compound of formula (II) in a solvent such as acetonitrileor dimethylformamide, tetrahydrofuran or dioxane and in the presence ofa base, for example sodium carbonate or potassium carbonate. Thereaction generally proceeds at room temperature to elevated temperatureto give a cyclic polyamine in which all amine nitrogens are protected.In general, a mixture of products will be obtained and we have foundthat the product can conveniently be purified by silica gelchromatography or crystallization.

[0029] Nucleophilic Attack of a Compound of Formula (III)

R⁵NH—(CR⁶R⁷)_(x)—R⁸  (III)

[0030] wherein R⁵ to R⁸ and x are as hereinbefore defined on the productof the reaction described at (I) above, and subsequently de-protectingthe amine nitrogens. The reaction with an excess of a compound offormula (III) is carried out under similar conditions to the reactionwith the polyamine V.

[0031] The de-protection step is suitably carried out by re-fluxing theprotected molecule in a mixture of aqueous HBr and acetic acid orconcentrated sulphuric acid, or in the case of diethoxyphosphoryl in thepresence of gaseous hydrogen chloride or gaseous hydrogen bromide inacetic acid; in the case of nitrobenzenesulfonyl deprotection, amercaptan such as thiophenol or mercaptoacetic acid in the presence of asuitable base such as potassium carbonate, cesium carbonate, sodiumhydroxide or lithium hydroxide in a solvent such as dimethylformamide,acetonitrile, tetrahydrofuran or dioxane is used. This reactiongenerally proceeds at room temperature to elevated temperatures to givea polyamine in which the nitrogens are de-protected. Alternatively, andaccordingly, a further aspect of the invention provides a process forthe preparation of compounds of formula (I) which comprises thefollowing steps:

[0032] (i) nucleophilic attack by the cyclic polyamine V having a singleunprotected amine nitrogen, all other amine nitrogens being protected,with an excess of a compound of formula (IV)

Y—CR¹R²—Ar—CR³R⁴—N(R⁵)—(CR⁶R⁷)_(x)—R⁸  (IV)

[0033] wherein R¹ to R⁴ and x, R⁶ to R⁸ and Ar are as hereinbeforedefined and Y is an active substituent which can be displaced by theunprotected nitrogen of cyclic polyamine V as hereinbefore defined. Inthis case, the intended substituent R⁵ is hydrogen but for convenience,the nitrogen is protected as a nitrobenzenesulfonyl ordiethoxyphosphoryl group.

[0034] The protected polyamine V is first reacted with a compound offormula (IV) using similar conditions to the reactions with compounds offormula (II) and formula (III) as described above and the product ofthis reaction is subjected to deprotection of the amine nitrogens on thepolyamine and at R⁵.

[0035] The deprotection steps were carried out as described above. Forconvenience, a sequential combination of these deprotection reactionsmay be used when a mixture of any of: methanesulphonyl;toluenesulphonyl; diethoxyphosphoryl; or nitrobenzenesulfonyl groups arepresent.

[0036] The novel compounds further comprise a macrocyclic compound ofgeneral formula (V):

V²—CR⁹R¹⁰—Ar²  (V)

[0037] where V² is a cyclic polyamine moiety having a total of 9 to 24members and from 3 to 6 optionally substituted amine nitrogens spaced bytwo or more optionally substituted carbon atoms from each other, andwhich may optionally comprise a fused aromatic or heteroaromatic; ring;where R⁹ and R¹⁰ may be the same or different and are independentlyselected from hydrogen or straight, branched or cyclic C₁₋₆ alkyl;further, where Ar² is an aromatic, fused aromatic, heterocyclic or fusedheterocyclic ring each optionally substituted at single or multiplepositions with electron-donating or withdrawing groups and/or aromaticand heterocyclic groups and their alkyl derivatives thereof; and theacid addition salts and metal complexes.

[0038] These novel compounds have demonstrated anti-HIV activity in anin vitro screen assay as presented in Table 1. These novel compoundshave also demonstrated biological activity in inhibiting CXCR-4 specificmonoclonal antibody (12G5) from binding to CXCR-4 on SUP-T1 cells by AMDcompounds. These data are shown in Table 2 for AMD3100(1,1′-[1,4-phenylenebis(methylene)]bis-1,4,8,11-tetraazacyclotetradecane,AMD3465(N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine)and six new compounds: AMD 7049; AMD 7050; AMD 7051; AMD 7058; AMD 7059;and AMD 7063.

[0039] Data showing inhibition of the increased Ca2+ flux induced bySDF-1 in SUP-T1 cells (inhibition of signal transduction) by AMDcompounds are shown in Table 3 for AMD3100, AMD3465 and compounds: AMD7049; AMD 7050; AMD 7051; AMD 7058; AMD 7059; and AMD 7063.

[0040] Several novel compounds also inhibited infection of the cell lineU87.CD4.CCR5 by the M-tropic HIV-1 strain BaL, which exclusivelyutilizes the CCR-5 co-receptor for entry. These data are shown in Table4.

[0041] The experimental procedures for the mAb binding assay, theinhibition of Ca²⁺ flux, and inhibition of infection by the HIV-1 BaLstrain in U87.CD4.CCR5 cells would be readily understood by the skilledartisan. For example, see: Schols, et al., J. Exp. Med. (1997)186:1383-1388; Schols, et al., Antiviral Research (1997) 35:147-156; andDonzella, et al., Nature Medicine (1998) 4:72-77. Also, thecharacterization of the CXCR-4 specific monoclonal antibody 12G5 istaught by Hoxie, et al., Cell (1996) 87:745-756.

[0042] Citation of the above documents is not intended as an admissionthat any of the foregoing is pertinent prior art. All statements as tothe date or representation as to the contents of these documents isbased on the information available to the applicants and does notconstitute any admission as to the correctness of the dates or contentsof these documents. Further, all documents referred to throughout thisapplication are incorporated in their entirety by reference herein.

[0043] Having now generally described the invention, the same will bemore readily understood through reference to the following exampleswhich are provided by way of illustration, and are not intended to belimiting of the present invention, unless specified.

[0044] As mentioned above, the compounds of the invention have activityagainst viral infections, especially retrovirus infections andspecifically HIV. Accordingly, a further aspect of the inventionprovides a compound of formula (I) or formula(V) for use in medicine.More specifically, there is provided the use of a compound of formula(I) or formula (V) in the manufacture of a medicament for the treatmentof HIV-infected patients. In the alternative, there is provided a methodof treating an HIV-infected patient comprising administering to saidpatient, a pharmaceutically effective amount of a compound of formula(I) or formula (V). Although compounds of formula (I) or formula (V)could be administered as the raw material, it is preferable to presentthem in the form of a pharmaceutical composition comprising a compoundof formula (I) or formula (V) as active ingredient in admixture with apharmaceutically acceptable diluent or carrier and optionally one ormore other therapeutic ingredients, such compositions providing afurther aspect of the invention.

[0045] In all aspects of the invention, it is understood that mesoforms, enantiomers and resolved optically active forms of the compoundsof formula (I) or formula (V) are also included. Also, it is to beconsidered within the invention, compounds of formula (I) or formula (V)diluted with non-toxic or other active substances.

BRIEF DESCRIPTION OF THE DRAWINGS

[0046]FIG. 1A, shows the structural formula of compound AMD 3465.

[0047]FIG. 1B, shows the structural formula of compound AMD 3538.

[0048]FIG. 1C; shows the structural formula of compound AMD 3500.

[0049]FIG. 1D; shows the structural formula of compound AMD 3499.

[0050]FIG. 1E; shows the structural formula of compound AMD 3498.

[0051]FIG. 1F; shows the structural formula of compound AMD 3497.

[0052]FIG. 1G; shows the structural formula of compound AMD 3516.

[0053]FIG. 1H; shows the structural formula of compound AMD 3530.

[0054]FIG. 11; shows the structural formula of compound AMD 3517.

[0055]FIG. 1J; shows the structural formula of compound AMD 3544.

[0056]FIG. 1K; shows the structural formula of compound AMD 3543.

[0057]FIG. 1L; shows the structural formula of compound AMD 3529.

[0058]FIG. 1M; shows the structural formula of compound AMD 7049.

[0059]FIG. 1N; shows the structural formula of compound AMD 7050.

[0060]FIG. 1O; shows the structural formula of compound AMD 7051.

[0061]FIG. 1P; shows the structural formula of compound AMD 7059.

[0062]FIG. 1Q; shows the structural formula of compound AMD 7063.

[0063]FIG. 1R; shows the structural formula of compound AMD 7058.

[0064]FIG. 1S; shows the structural formula of compound AMD 7032.

[0065]FIG. 1T; shows the structural formula of compound AMD 7048.

[0066]FIG. 1U; shows the structural formula of compound AMD 7060.

[0067]FIG. 1V; shows the structural formula of compound AMD 7061.

[0068]FIG. 1W; shows the structural formula of compound AMD 3451.

[0069]FIG. 1X; shows the structural formula of compound AMD 3454.

[0070]FIG. 1Y; shows the structural formula of compound AMD 3472.

[0071]FIG. 1Z; shows the structural formula of compound AMD 3526.

[0072]FIG. 1AA; shows the structural formula of compound AMD 3100.

[0073]FIG. 1AB; shows the structural formula of compound AMD 3484.

MODES OF CARRYING OUT THE INVENTION

[0074] Terms as used herein are based upon their art recognized meaningunless otherwise indicated and should be clearly understood by theordinary skilled artisan. The present invention will now be illustratedby the following preparative Examples.

General Procedure A1-[1-Methylene-4-(bromomethylene)phenylene]-4,8,11-tris(diethoxyphosphoryl)-1,4,8,11-tetraazacyclotetradecane

[0075] To a stirred solution of4,8,11-Tris(diethoxyphosphoryl)-1,4,8,11-tetraazacyclotetradecane (seeBridger, et al., J Med. Chem. (1995) 38:366-378) (6.1 g, 0.01 mol) andK₂CO₃ (1.89 g, 0.013 mol) in CH₃CN (150 ml) was addedα,α′-dibromo-p-xylene (13.2 g, 0.05 mol) and the reaction mixturestirred at 70° C. for 1 hour. The solution was cooled to roomtemperature and the solvent removed under reduced pressure. The residuewas partitioned between brine (50 ml) and CH₂Cl₂ (100 ml). The organicphase was separated, dried (Na₂SO₄) and concentrated to a minimumvolume. The solid was filtered off and the solvent evaporated underreduced pressure to give the crude product as a pale yellow oil.Purification by column chromatography in silica gel (CH₂Cl₂\CH₃OH, 25:1)gave1-[1-methylene-4-(bromo-methylene)phenylene]-4,8,11-tris(diethoxyphosphoryl-1,4,8,11-tetraazacyclotetra-decane(4.7 g, 59%) as a pale yellow oil. ¹H NMR (CDCl₃) δ 1.21-1.37 (m, 18H),1.66-1.74 (m, 2H), 1.82-1.91 (m, 2H), 2.30-2.35 (m, 2H), 2.58-2.63 (m,2H), 2.99-3.16 (m, 12H), 3.48 (s, 2H), 3.95-4.07 (m, 12H), 4.48 (s, 2H),7.21-7.35 (4H).

General Procedure B Second Alkylation of the Bromobenzyl CyclamIntermediate with an Amine (See for Example: Bridger, et al., J. Med.Chem. (1995) 38:366-378)

[0076] To a solution of the appropriate amine (5.0 equiv.) in dry CH₃CN(5 mL) containing a suspension of K₂CO₃ (1.5 equiv.) at 80° C. was addeddropwise with stirring a solution of1-[1-methylene-4-(bromomethylene)phenylene]-4,8,11-tris(diethoxyphosphoryl-1,4,8,11-tetraazacyclotetradecane(0.6 mmol) in CH₃CN (10 ml) over 15-20 min. After stirring for a further1 hour at 80° C. the solution was concentrated to dryness and theresidue was partitioned between CH₂Cl₂ and water. The organic layer wasseparated and washed with water (3×) then dried (MgSO₄) and evaporated.The crude residue was purified by column chromatography on silica geleluting with 5-15% MeOH/CH₂Cl₂ to afford a viscous oil.

General Procedure C De-Protection of the Diethoxyphosphoramidate Groupsusing HBr/HOAc at Room Temperature (See for Example: Bridger. et al., J.Med. Chem. (1995) 38:366-378)

[0077] To a stirred solution of the protected cyclam derivative fromprocedure B (0.1-0.5 mmol) in acetic acid (3 mL) was added 30% HBr inacetic acid (Aldrich, 5 mL) and the solution was stirred at roomtemperature for 14 hours. The resulting precipitate was collected byfiltration and washed with acetic acid then Et₂O. The solid was thendissolved in H₂O (3 mL) and treated with charcoal (100 mg) and themixture was heated to 80° C. for 30 min. The hot solution was filteredthrough celite and the filtrate was concentrated to approximately 1 mLafter which acetic acid was added resulting in the immediate formationof a white precipitate. The white solid was collected by filtration anddried in vacuo.

[0078] The following compounds were prepared by these methods:

EXAMPLE 1N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(amino-methyl)pyridinehexahydrobromide (AMD 3465)

[0079] White solid: Mp 200-205° C. (dec); ¹H NMR (D₂O) δ 2.04 (m, 4H),3.20-3.40 (m, 8H), 3.40-3.60 (m, 8H), 4.34 (s, 2H), 4.38 (s, 2H), 4.51(s, 2H), 7.50 (m, 4H), 7.75 (t, 1H, J=6.6 Hz), 7.82 (d, 1H, J=7.9 Hz),8.26 (t, 1H, J=7.9 Hz), 8.63 (d, 1H, J=5.3 Hz); ¹³C NMR (D₂O) δ 18.30,18.96, 37.04, 37.28, 37.40, 40.92, 41.13, 41.49, 44.26, 47.61, 48.01,51.29, 58.88, 127.46, 127.75, 130.40, 131.05, 131.23, 131.47, 132.10,132.44, 144.95, 145.81, 146.01; FAB MS m/z 493 (M+H⁸¹Br, 7), 491(M+H⁷⁹Br, 7), 411 (M+H, 100).

[0080] Anal. (C₂₄H₃₈N₆.6HBr); Calc. C, 32.36; H, 4.98; N, 9.44; Br,53.21. Found C, 32.20; H, 5.00; N, 9.30; Br, 53.10.

EXAMPLE 2N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene]-N-methyl-2-(aminomethyl)pyridinehexahydrobromide hydrate (AMD 3538)

[0081] White solid: Mp 220-225° C. (dec); ¹H NMR (D₂O) δ 2.06 (m, 4H),2.76 (s, 3H); 3.20-3.65 (m, 16H), 4.47 (bs, 4H), 4.65 (s, 2H), 7.54 (bs,4H), 7.80 (t, 1H), 7.87 (d, 1H), 8.28 (t, 1H), 8.68 (d, 1H); ¹³C NMR(D₂O) δ 18.14, 18.75, 18.89, 36.74, 37.04, 37.15, 37.62, 40.38, 40.72,40.91, 41.28, 44.05, 47.50, 56.98, 58.88, 60.28, 127.60, 128.86, 130.78,130.96, 132.16, 132.64, 144.91, 145.04, 146.12; FAB MS m/z 507 (M+H⁸¹Br,27), 507 (M+H⁷⁹Br, 22), 425 (M+H, 100).

[0082] Anal. (C₂₅H₄₀N₆.6HBr.1.5H₂O); Calc. C, 32.04; H, 5.27; N, 8.97;Br, 51.16. Found C, 31.88; H, 5.30; N, 8.93; Br, 51.00.

EXAMPLE 3N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-4-)amino-methyl)pyridinehexahydrobromide (AMD 3500)

[0083] White solid: mp 201-204° C. (dec); ¹H NMR (D₂O) δ 1.91-2.12 (m,4H), 3.00-3.49 (m, 16H), 4.13 (s, 2H), 4.34 (s, 2H), 4.53 (s, 2H),7.39-7.57 (m, 4H), 8.02 (d, 2H, J=6.3 Hz), 8.74 (d, 2H, J=6.3 Hz); ¹³CNMR (D₂O) δ 18.26, 18.88, 36.94, 37.29, 37.36, 40.89, 41.06, 41.44,44.21, 47.61, 49.17, 51.43, 59.02, 127.84, 130.21, 131.64, 132.15,132.45, 142.19, 151.67; FAB MS m/z 493 (M+H⁸¹Br, 8), 491 (M+H⁷⁹Br, 10),411 (M+H, 83), 320 (37), 247 (58), 201 (100).

[0084] Anal. (C₂₄H₃₈N₆.6HBr); Calc. C, 32.17; H, 4.95; N, 9.34; Br,53.50. Found C, 32.16; H, 5.03; N, 9.41; Br, 53.28.

EXAMPLE 4N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene]-3-(amino-methyl)pyridinehexahydrobromide (AMD 3499)

[0085] White solid: mp 198-202° C. (dec); ¹H NMR (D₂O) δ 1.83-2.07 (m,4H), 2.96-3.47 (m, 16H), 4.11 (s, 2H), 4.32 (s, 2H), 4.49 (s, 2H),7.38-7.56 (m, 4H), 8.04 (t, 1H, J=6.4 Hz), 8.63 (d, 1H, J=8.3 Hz), 8.76(d, 1H, J=5.6 Hz), 8.86 (s, 1H); ¹³C NMR (D₂O) δ 18.23, 18.87, 36.92,37.29 (2C), 40.88, 41.05, 41.43, 44.17, 47.22, 47.60, 51.18, 59.04,128.29, 130.01, 131.49, 132.14, 132.66 (2C), 142.55, 142.76, 148.98; FABMS m/z 493 (M+H⁸¹Br, 7), 491 (M+H⁷⁹Br, 6), 411 (M+H, 100), 320 (33), 247(24).

[0086] Anal. (C₂₄H₃₈N₆.6HBr); Calc. C, 32.17; H, 4.95; N, 9.34; Br,53.50. Found C, 32.08; H, 5.02; N, 9.25; Br, 53.28.

EXAMPLE 5N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-(2-amino-methyl-5-methyl)pyrazinepentahydrobromide (AMD 3498)

[0087] White solid: mp 194-197° C. (dec); ¹H NMR (D₂O) δ 1.93-2.12 (m,4H), 2.42 (s, 3H), 3.25 (s, 8H), 3.48 (s, 8H), 4.28 (s, 2H), 4.30 (s,2H), 4.33 (s, 2H), 7.44 (s, 4H), 8.33 (s, 1H), 8.46 (s, 1H); ¹³C NMR(D₂O) δ 18.01, 18.72, 19.80, 36.66, 37.05, 37.13, 40.70, 40.89, 41.27,43.99, 47.47, 48.14, 50.61, 59.06, 129.97, 131.43, 132.04, 132.99,140.93, 144.98, 146.49, 153.51; FAB MS m/z 509 (M+H⁸¹Br, 17), 507(M+H⁷⁹Br, 15), 426 (M+H, 100), 320 (21), 247 (20).

[0088] Anal. (C₂₄H₃₉N₇.5.5HBr); Calc. C, 33.10; H, 5.15; N, 11.26; Br,50.47. Found C, 32.80; H, 5.41; N, 11.00; Br, 50.58.

EXAMPLE 6N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(amino-ethyl)pyridinehexahydrobromide (AMD 3497)

[0089] White solid: mp 195-198° C. (dec); ¹H NMR (D₂O) δ 1.98-2.17 (m,4H), 3.20-3.38 (m, 8H), 3.38-3.63 (m, 12H), 4.27 (s, 2H), 4.39 (s, 2H),7.50 (s, 4H), 7.80-7.89 (m, 2H), 8.42 (m, 1H), 8.58 (d, 1H, J=5.8 Hz);¹³C NMR (D₂O) δ 18.51, 19.14, 29.85, 37.56 (3C), 41.21, 41.41, 41.82,44.57, 45.27, 47.83, 51.10, 58.74, 126.35, 127.93, 130.66, 131.27,131.99, 132.69, 141.89, 147.79, 150.91; FAB MS m/z 507 (M+H⁸¹Br, 40),505 (M+H ⁷⁹Br, 34), 425 (M+H, 100).

[0090] Anal. (C₂₅H₄₀N₆.6HBr); Calc. C, 32.99; H, 5.09; N, 9.23; Br,52.67. Found C, 32.79; H, 5.34; N, 9.11; Br, 52.45.

EXAMPLE 7N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(amino-methyl)thiophenepentahydrobromide (AMD 3516)

[0091] White solid: mp 245-248° C. (dec); ¹H NMR (D₂O) δ 1.87-2.12 (m,4H), 3.02-3.51 (m, 16H), 4.17 (s, 4H), 4.38 (s, 2H), 6.97 (t, 1H, J=3.9Hz), 7.13 (d, 1H, J=3.1 Hz), 7.41 (s, 5H); ¹³C NMR (D₂O) δ 18.80, 19.52,38.03, (3C), 41.59 (2C), 42.21, 44.89 (2C), 48.15, 49.83, 58.52, 128.13,129.12, 131.15, 131.47, 131.50, 131.90, 132.42, 132.87; FAB MS m/z 498(M+H⁸¹Br, 11), 496 (M+H⁷⁹Br, 9), 416 (M+H, 53), 218 (100), 201 (64).

[0092] Anal. (C₂₃H₃₇N₅S.5HBr); Calc. C, 33.68; H, 5.16; N, 8.54; Br,48.71. Found C, 33.85; H, 5.22; N, 8.50; Br, 48.52.

EXAMPLE 8N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(amino-ethyl)mercaptanpentahydrobromide dihydrate (AMD 3530)

[0093] White solid: mp 234-236° C. (dec); ¹H NMR (D₂O) δ 1.75-2.05 (m,4H), 2.75-3.45 (m, 20H), 4.05 (s, 2H), 4.15 (s, 2H), 7.35 (s, 4H); FABMS m/z 462 (MH+H⁸¹Br, 15), 460 (MH+H⁷⁹Br, 15), 380 (M+H, 100), 300 (64),279 (47), 239 (49).

[0094] Anal. (C₂₀H₃₇N₅S.5HBr.2H₂O.0.5HOAc) requires C, 29.67; H, 5.69;N, 8.24; Br, 46.99. Found C, 29.31; H, 5.72; N, 8.25; Br, 46.64.

EXAMPLE 9N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-amino-benzylaminepentahydrobromide (AMD 3517)

[0095] White solid: mp 203-206° C. (dec); ¹H NMR (D₂O) δ 1.85-2.13 (m,4H), 3.02-3.58 (m, 16H), 4.23 (s, 2H), 4.31 (s, 4H), 7.23-7.54 (m, 8H);¹³C NMR (D₂O) δ 18.03, 19.29, 37.78 (3C), 41.37 (2C), 42.00, 44.82,46.25, 47.96, 51.16, 58.68, 124.04, 124.40, 129.40, 130.75, 131.21 (2C),131.88, 131.96, 132.46, 132.83; FAB MS m/z 507 (M+H⁸¹Br, 15), 505(M+H⁷⁹Br, 18), 425 (M+H, 100), 320 (30), 201 (51).

[0096] Anal. (C₂₅H₄₀N₆.5.75HBr.0.5H₂O). Calc. C, 33.42; H, 5.19; N,9.35; Br, 51.14. Found C, 33.69; H, 5.35; N, 9.00; Br, 51.13.

EXAMPLE 10N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-4-amino-benzylaminehexahydrobromide (AMD 3544)

[0097] Yellow solid: mp 120-125° C. (dec); ¹H NMR (D₂O) δ 1.8-2.0 (m,4H), 2.9-3.4 (m, 16H), 4.1 (s, 2H), 4.18 (s, 4H), 7.2-7.5 (m, 8H); ¹³CNMR (D₂O) δ 18.86, 19.57, 38.14, 41.76, 43.74, 45.14, 48.24, 50.14,50.42, 51.49, 58.38, 124.13, 131.13, 131.30, 131.83, 131.92, 131.96,132.67; FAB MS m/z 507 (M+H⁸¹Br, 5), 505 (M+H⁷⁹Br, 5), 425 (M+H, 45),201 (47), 155 (75), 106 (100).

[0098] Anal. (C₂₅H₄₀N₆.6HBr.HOAc) requires C, 33.43; H, 5.19; N, 8.66;Br, 49.42; O, 3.30. Found C, 33.42; H, 5.49; N, 8.62; Br, 49.23.

EXAMPLE 11N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-4-(amino-ethyl)imidazolehexahydrobromide (AMD 3543)

[0099] Off white solid: mp 135-140° C. (dec); ¹H NMR (D₂O) δ 1.75 (m,2H), 190 (m, 2H), 2.70-3.27 (m, 20H), 3.77 (s, 2H), 4.14 (s, 2H), 7.18(s, 1H), 7.25 (d, 2H, J=7.97 Hz), 7.37 (d, 2H, J=7.97 Hz), 8.48 (s, 1H);FAB MS m/z 496 (M+H⁸¹Br, 5), 494 (M+H⁷⁹Br, 5), 414 (M+H, 17), 201 (15).

[0100] Anal. (C₂₃H₃₉N₇.6HBr) requires C, 30.73; H, 5.04; N, 10.91; Br,53.32. Found C, 30.39; H, 5.41; N, 10.41; Br, 53.66.

EXAMPLE 12N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-benzylaminepentahydrobromide (AMD 3529)

[0101] Off white solid: mp 245-250° C. (dec); ¹H NMR (D₂O) δ 1.9-2.1 (m,4H), 3.2-3.6 (m, 16H), 4.12 (s, 2H), 4.15 (s, 2H), 4.36 (s, 2H), 7.30(s, 5H), 7.41 (d, 2H, J=8.3 Hz) 7.46 (d, 2H, J=8.3 Hz); ¹³C NMR (D₂O) δ18.43, 19.06, 37.29, 37.46, 37.63, 41.09, 41.32, 41.68, 44.46, 47.74,50.18, 51.00, 58.79, 129.53, 129.97, 130.18, 130.35, 130.68, 131.18,131.92, 133.14; FAB MS m/z 492 (M+H⁸¹Br, 13), 490 (M+H⁷⁹Br, 13), 410(M+H, 100), 201 (36).

[0102] Anal. (C₂₅H₃₉N₅.5HBr); requires C, 36.88; H, 5.45; N. 8.60; Br,49.07. Found C, 36.79; H, 5.56; N, 8.48; Br, 48.79.

[0103] The compounds of the invention were tested in a screen by the MTTmethod (J. Virol. Methods (1988) 120:309-321). MT-4 cells (2.5×10⁴/well)were challenged with HIV-1 (HTLV-IIIB) or HIV-2 (LAV-2 ROD) at aconcentration of 100 CCID₅₀ and incubated in the presence of variousconcentrations of the test compounds, which were added immediately afterchallenge with the virus. After 5 days culture at 37° C. in a CO₂incubator, the number of viable cells was assessed by the MTT(tetrazolium) method. Antiviral activity and cytotoxicity of thecompounds are expressed in Table 1 below as EC₅₀ (μg/ml) and CC₅₀(μg/ml), respectively. The potential therapeutic usefulness was assessedby calculating a Selectivity Index (SI) corresponding to the ratio ofCC₅₀ to EC₅₀. TABLE 1 Anti-HIV activity data EC₅₀ (μg/mL) Compound CC₅₀(μg/mL) HIV-1 (IIIB) HIV-2 SI HIV-1  1 AMD3465 >250 0.008 0.032   3 ×10⁴  2 AMD3538 209 0.1 6.7 2.0 × 10³  3 AMD3500 >250 0.6 10.3 417  4AMD3499 >250 1.8 28.5 138  5 AMD3498 >250 0.2 7.1 1.2 × 10³  6AMD3497 >250 1.8 3.8 138  7 AMD3516 158 0.7 9.8 225  8 AMD3530 175 0.52.0 350  9 AMD3517 153 0.8 10.6 191 10 AMD3544 222 0.7 3.7 317 11AMD3543 239 0.2 1.0   1 × 10³ 12 AMD3529 130 0.4 2.6 325

[0104] In this field of study, it is considered that any compoundexhibiting a Selectivity Index of greater than 100 has the considerablepotential for further study. HIV is one of the most challenging virusesto combat, and the results given above provide an indication of activityagainst other retroviruses and against other viruses in general.

EXAMPLE 13N-[4-(1,4,7-Triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7049)

[0105] N,N′-Bis(2-Nitrobenzenesulfonyl)-1,7-heptanediamine

[0106] To a stirred solution of 1,7-heptanediamine (5.01 g, 38.5 mmol)and Et₃N (13.5 mL, 96.9 mmol) in CH₂Cl₂ (70 mL) was added a solution of2-nitrobenzenesulfonyl chloride (18.80 g, 84.83 mmol) in CH₂Cl₂ (40 mL).The mixture was stirred at room temperature under nitrogen for 72 h andthen concentrated in vacuo. The residue was stirred in diethyl ether(100 mL), and the precipitate was collected by filtration and washedwith H₂O (300 mL) followed by diethyl ether (300 mL) to give a graysolid (18.5 g, 96%): ¹H NMR (DMF-d₇) δ 1.21 (m, 6H), 1.49 (m, 4H), 3.04(m, 4H), 7.87 (m, 2H), 7.95 (m, 4H), 8.04 (m, 2H), 8.15 (m, 2H).

General Procedure D4-Diethoxyphosphoryl-1,7-bis(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetradecane

[0107] To a stirred solution ofN,N′-bis(2-nitrobenzenesulfonyl)-1,7-heptanediamine (9.00 g, 18.0 mmol)and Cs₂CO₃ (17.8 g, 54.6 mmol) in DMF (500 mL) under nitrogen maintainedat 80° C. was added dropwise a solution ofN-(diethoxyphosphoryl)-O,O′-bis(2-methylsulfonyl)di-ethanolamine(Bridger, et al., J. Med. Chem. (1995) 38:366-378) (7.95 g, 20.0 mmol)in DMF (50 mL) over 8 h. Heating was continued for a further 17 h andthe mixture was then allowed to cool and concentrated in vacuo. Theresidue was partitioned between CHCl₃ (140 mL) and H₂O (80 mL) and theaqueous layer was separated and extracted with CHCl₃ (3×40 mL). Thecombined organic extracts were dried (MgSO₄) and concentrated in vacuoand the residue was purified by column chromatography on silica gel(ethylacetate) to give the desired macrocycle as a yellow crystallinesolid (2.85 g, contaminated with DMF).

[0108] To remove the unwanted DMF impurity, the residue was dissolved inEtOAc (75 mL), and the solution was washed sequentially with 5% NaHCO₃(2×10 mL) and brine (5×10 mL), dried (MgSO₄) and evaporated to give ayellow amorphous solid (2.52 g, 20%): ¹H NMR (CDCl₃) δ 1.32 (t, 6H,J=7.1 Hz), 1.51 (m, 6H), 1.61 (m, 4H), 3.33 (m, 12H), 4.03 (m, 4H), 7.61(m, 2H), 7.71 (m, 4H), 8.03 (m, 2H).

General Procedure E Synthesis of1,7-Bis(2-nitrobenzenesulfonyl-1,4,7-triazacyclotetradecane

[0109] To a stirred suspension of the macrocycle from above (1.88 g,2.66 mmol) in acetic acid (5 mL) was added a freshly prepared solutionof saturated HBr(g) in acetic acid (20 mL) and the resulting homogeneoussolution was stirred at room temperature for a further 22 h. Addition ofdiethyl ether (250 mL) to the reaction mixture gave a precipitate thatwas allowed to settle to the bottom of the flask and the supernatantsolution was decanted. The precipitate was washed with ether bydecantation (repeated 3×) and the residue was then partitioned betweenCH₂Cl₂ (40 mL) and 1N aqueous NaOH (25 mL). The separated aqueous layerwas extracted with CH₂Cl₂ (2×20 mL) and the combined organic extractswere washed with brine (20 mL), then dried (MgSO₄) and concentrated invacuo to give a yellow amorphous solid (1.23 g, 81%): ¹H NMR (CDCl₃) δ1.46-1.67 (m, 10H), 2.90 (m, 4H), 3.34 (m, 8H), 7.61 (m, 2H), 7.70 (m,4H), 7.97 (m, 2H).

[0110] 4-Bromomethylbenzyl alcohol

[0111] To a solution of methyl 4-bromomethylbenzoate (5.73 g, 25 mmol)in dry CH₂Cl₂ (150 mL) cooled to −78° C. with stirring under nitrogenwas added dropwise a solution of DIBAL-H (82.5 mL, 1.0 M solution inTHF). Stirring was continued for 1.5 h at −78° C., and the reactionmixture was then allowed to warm to 0° C. and quenched with H₂O. Theorganic layer was separated and the aqueous was extracted with CH₂Cl₂(2×100 mL). The combined organic extracts were dried (MgSO₄) andevaporated to give the desired alcohol (5.0 g, 100%) as a white solid:¹H NMR (CDCl₃) δ 1.84 (br, 1H), 4.49 (s, 2H), 4.67 (s, 2H), 7.33 (d, 2H,J=8.2 Hz), 7.38 (d, 2H, J=8.2 Hz).

[0112] N-(2-Nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0113] A solution of 2-nitrobenzenesulfonylchloride (16.62 g, 0.075 mol)in dry CH₂Cl₂ (120 mL) was added dropwise via cannula to a stirredsolution of 2-(aminomethyl)pyridine (5.41 g, 0.05 mol) and Et₃N (13.9mL, 0.10 mol) in dry CH₂Cl₂ (150 mL) under nitrogen. The reactionmixture was stirred for three hours at room temperature, and thenquenched with water (20 mL). The aqueous layer was separated andextracted with EtOAc (5×80 mL). The combined organic extracts were dried(MgSO₄) and evaporated to small volume to give a white precipitate whichwas collected by filtration and washed with cold CH₂Cl₂ to give thedesired product (11.37 g, 78%) as a white solid: ¹H NMR (Acetone-d₆) δ4.46 (s, 2H), 7.19 (dd, 1H, J=7.4, 4.5 Hz), 7.25-7.35 (br s, 1H), 7.39(d, 1H, J=7.7 Hz), 7.68 (ddd, 1H, J=7.7, 7.5, 1.8 Hz), 7.76-7.88 (m,2H), 7.94 (dd, 1H, J=7.7, 1.5 Hz), 8.04 (dd, 1H, J=7.5, 1.8 Hz), 8.38(d, 1H, J=4.5 Hz).

[0114]N-[1-Methylene-4-(hydroxymethylene)phenylene]-N-(2-Nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0115] A mixture of N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(5.87 g, 20 mmol), 4-bromomethylbenzyl alcohol (4.02 g, 20 mmol) andK₂CO₃ (5.53 g, 40 mmol) in dry CH₃CN (150 mL) were heated at 60° C. for4 h with stirring under nitrogen. The mixture was then allowed to coolto room temperature, the solvent evaporated and the residue waspartitioned between water and CH₂Cl₂. The separated aqueous phase wasextracted with CH₂Cl₂, and the combined organic extracts were dried(MgSO₄) and evaporated. The residue was suspended in ethylacetate/hexane (1:1) and collected by filtration to give the desiredproduct (6.87 g, 83%) as a white solid: ¹H NMR (CDCl₃) δ 1.78 (t, 1H,J=5.8 Hz), 4.58 (s, 2H) 4.60 (s, 2H), 4.64 (d, 2H, J=5.8 Hz), 7.13-7.26(m, 6H), 7.54-7.59 (m, 2H), 7.66-7.68 (m, 2H), 7.98 (d, 1H, J=7.4 Hz),8.40 (d, 1H, J=3.8 Hz).

[0116]N-[1-Methylene-4-(chloromethylene)phenylene]-N-(2-Nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0117] To a stirred solution of the alcohol from above (1.91 g, 4.62mmol) and Et₃N (2.0 mL, 14 mmol) in CH₂Cl₂ (20 mL) cooled in an ice bathunder nitrogen, was added methanesulfonyl chloride (0.73 mL, 9.4 mmol)and the reaction mixture was then heated to reflux for a further 6 h.The solution was diluted with CH₂Cl₂ (60 mL) and washed with 10% aqueousHCl (2×20 mL), 5% aqueous NaHCO₃ (20 mL), and H₂O (25 mL) then dried(MgSO₄) and concentrated in vacuo to give an orange oil (1.95 g, 98%):¹H NMR (CDCl₃) δ 4.52 (s, 2H), 4.60 (s, 4H), 7.12-7.26 (m, 6H), 7.55 (m,2H), 7.67 (d, 2H, J=4.0 Hz), 7.94 (d, 1H, J=8.0 Hz), 8.41 (d, 1H, J=4.8Hz). This was used without further purification.

General Procedure FN-[4-[1,7-Bis(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetra-decanyl]-1,4-phenylenebis(methylene)]-N-(2-Nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0118] A mixture of1,7-bis(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetradecane (1.1 g, 1.9mmol), the chloride from above (0.98 g, 2.3 mmol) and K₂CO₃ (0.85 g, 6.2mmol) were heated to reflux in CH₃CN (30 mL) under nitrogen for 62 h.The solvent was evaporated in vacuo and the residue was partitionedbetween CH₂Cl₂ (100 mL) and brine (70 mL). The aqueous phase wasseparated and extracted with CH₂Cl₂ (40 mL) and the combined organicextracts were dried (MgSO₄) and concentrated in vacuo. The residue waspurified by column chromatography on silica gel (3% MeOH/CH₂Cl₂) and theevaporated fractions containing the desired product were subjected to asecond column purification on silica gel (ethyl acetate) to give a paleyellow amorphous solid (940 mg, 49%): ¹H NMR (CDCl₃) δ 1.44 (br s, 6H),1.60 (br s, 4H), 2.75 (m, 4H.), 3.23-3.33 (m, 8H), 3.59 (s, 2H), 4.58(s, 2H), 4.59 (s, 2H), 7.08-7.20 (m, 6H), 7.55-7.70 (m, 10H), 7.82 (dd,2H, J=7.6, 1.6 Hz), 7.99 (d, 1H, J=7.8 Hz), 8.40 (d, 1H, J=4.7 Hz).

[0119]N-[4-(1,4,7-Triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridinePentahydrobromide Dihydrate

[0120] The intermediate from above (870 mg, 0.90 mmol), K₂CO₃ (1.15 g,8.32 mmol), and thiophenol (0.33 mL, 3.2 mmol) were stirred in DMF (12mL) for 7.5 h at room temperature. The mixture was concentrated in vacuoand the residue was partitioned between CH₂Cl₂ (30 mL) and H₂O (15 mL).The organic phase was separated, washed with 5% NaHCO₃ (10 mL) then H₂O(10 mL) then dried (MgSO₄) and concentrated in vacuo. The yellow residuewas purified by column chromatography on basic alumina (CH₂Cl₂, 1%MeOH/CH₂Cl₂, and 10% MeOH/CH₂Cl₂)to give the free base as a yellow oil(134 mg, 36%): ¹H NMR (CDCl₃) δ 1.48 (br s, 6H), 1.60 (br s, 4H), 2.61(m, 12H), 3.56 (s, 2H), 3.83 (s, 2H), 3.92 (s, 2H), 7.16 (m, 1H), 7.24(m, 2H), 7.32 (m, 3H), 7.79 (m, 1H), 8.56 (d, 1H, J=4.7 Hz).

[0121] The free base (134 mg, 0.33 mmol) was dissolved in EtOH (4 mL)and a freshly prepared solution of saturated HBr(g) in EtOH (9 mL) wasadded, giving a white precipitate. The mixture was stirred for 5 min anddiethyl ether (15 mL) was added. The precipitate was allowed to settleto the bottom of the flask and the supernatant solution was decanted.The solid was then dissolved in MeOH (5 mL) and re-precipitated with alarge volume of ether, washed with ether by decantation (15×) andfinally, the last traces of ether were removed by evaporation at reducedpressure (room temperature). Drying, the solid in vacuo at 40° C. for 16h, gave the desired product as a white solid (178 mg, 63%): ¹H NMR(DMSO-d₆) δ 1.44 (br s, 6H), 1.75 (br s, 4H), 3.04 (br s, 8H), 3.37 (m,4H), 4.06 (br s, 2H), 4.31 (s, 211), 4.38 (s, 2H), 7.52-7.68 (m, 6H),8.01 (m, 1H), 8.70 (d, 1H, J=5.0 Hz); FAB-MS m/z 492 (MH+H⁸¹Br), 490(MH+H⁷⁹Br), 410 (M+H). Anal. Calcd for C₂₅H₃₉N₅.5HBr.0.1Et₂O.2.3H₂O: C,35.35; H, 5.79; N, 8.11; Br, 46.29. Found: C, 35.55; H, 5.70; N, 8.18;Br, 46.17.

EXAMPLE 14N-[7-(4,7,10,17-Tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7050)

[0122] 2,6-Bis(2-aminoethyl)pyridine was prepared as described inBridger, et al., U.S. Pat. No. 5,698,546, which is hereby incorporatedin its entirety by reference herein.

[0123] 2,6-Bis[N-(2-nitrobenzenesulfonyl)-2-aminoethyl]pyridine

[0124] To a stirred solution of 2,6-Bis(2-aminoethyl)pyridine (2.7 g, 16mmol) and Et₃N (5.7 mL, 41 mmol) in CH₂Cl₂ (35 mL) was added2-nitrobenzenesulfonyl chloride (8.01 g, 36.1 mmol) in CH₂Cl₂ (20 mL)and the mixture was stirred at room temperature under nitrogen for 42 h.The mixture was washed with brine (25 mL) and the organic phase wasdried (MgSO₄) and concentrated in vacuo. The brown residue was purifiedby column chromatography on silica gel (50% then 60% THF/hexane) to givea pale yellow solid (5.2 g, 59%): ¹H NMR (CDCl₃) δ 3.01 (m, 4H), 3.52(m, 4H), 6.38 (m, 2H), 6.94 (d, 2H, J=7.7, Hz), 7.47 (t, 1H, J=7.7 Hz),7.72 (m, 4H), 7.82 (m, 2H), 8.13 (m, 2H).

[0125]7-Diethoxyphosphoryl-4,10-Bis(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene

[0126] Using General Procedure D: Reaction of2,6-bis[N-(2-nitrobenzenesulfonyl)-2-aminoethyl]pyridine (5.2 g, 9.7mmol) andN-(diethoxyphosphoryl)-O,O′-bis(2-methylsulfonyl)di-ethanolamine (4.25g, 10.7 mmol) followed by silica gel column purification (60% then 90%THF/hexane) of the reaction products gave the title compound as a yellowamorphous solid (1.48 g, 21%): ¹H NMR (CDCl₃) δ 1.23 (t, 6H, J=7.1 Hz),2.60 (m, 4H), 2.98-3.08 (m, 8H), 3.84-3.94 (m, 8H), 7.11 (d, 2H, J=7.6Hz), 7.56-7.74 (m, 7H), 8.07 (m, 2H).

[0127]4,10-Bis(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene

[0128] Using General Procedure E: Reaction of7-diethoxyphosphoryl-4,10-bis(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene(1.04 g, 1.4 mmol) gave the title compound as a yellow amorphous solid(744 mg, 88%): ¹H NMR (CDCl₃) δ 2.81 (m, 4H), 3.08 (m, 4H), 3.33 (m,4H), 3.88 (m, 4H), 7.07 (d, 2H, J=7.7 Hz), 7.54-7.71 (m, 711, 8.02 (m,2H).

[0129]N-[7-[4,10-Bis(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0130] Using General Procedure F: Reaction of4,10-bis(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene(740 mg, 1.2 mmol) andN-[1-methylene-4-(chloromethylene)phenylene]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(610 mg, 1.4 mmol) followed by silica gel column purification (50% then80% THF/hexane) of the reaction products gave the title compound as ayellow amorphous solid (648 mg, 54%): ¹H NMR (CDCl₃) δ 2.26 (m, 4H),3.03 (m, 8H), 3.37 (s, 2H), 3.94 (m, 4H), 4.56 (s, 2H), 4.57 (s, 2H),6.95-7.17 (m, 8H), 7.52-7.72 (m, 11H), 7.85 (m, 2H), 7.98 (d, 1H, J=7.7Hz), 8.39 (d, 1H, J=4.8 Hz).

General Procedure GN-[7-(4,7,10,17-Tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridineHexahydrobromide Trihydrate

[0131] To a solution ofN-[7-[4,10-bis(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(640 mg, 0.64 mmol in DMF (9 mL) containing K₂CO₃ (806 mg, 5.83 mmol)was added thiophenol (0.24 mL, 2.3 mmol) and the mixture was stirred atroom temperature for 2 h. The mixture was concentrated in vacuo and theresidue partitioned between ethyl acetate (30 mL) and water (10 mL). Theorganic phase was separated and extracted with 5% NaHCO₃ (3×5 mL) thenbrine (5 mL; The combined aqueous, phases were extracted with CH₂Cl₂(3×10 mL). The combined organic extracts were dried (MgSO₄) andevaporated and the residue was purified by column chromatography onalumina (CH₂Cl₂ followed by 10% MeOH/CH₂Cl₂) to give the free base ofthe title compound as a yellow oil (83 mg, 29%): ¹H NMR (CDCl₃) δ 2.57(m, 8H), 3.01 (s, 8H), 3.36 (s, 2H), 3.78 (s, 2H), 3.92 (s, 2H), 6.64(d, 2H, J=8.0 Hz), 7.07 (m, 4H), 7.18 (m, 1H), 7.33 (d, 1H, J=7.7 Hz),7.67 (m, 2H), 8.58 (d, 1H, J=4.8 Hz).

[0132] The free base (74 mg, 0.17 mmol) was dissolved in MeOH (3 mL) anda freshly prepared solution of saturated HBr(g) in MeOH (7 mL) was addedgiving a white precipitate. The mixture was stirred for 5 min anddiethyl ether was added (10 mL), the solid was allowed to settle to thebottom of the flask and the supernatant solution decanted. The solid waswashed by decantation with MeOH (5×5 mL) then ether (10×5 mL) and thelast traces of ether were removed by evaporation in vacuo followed bydrying in vacuo at 40° C. for 17.5 h to give the title compound as awhite solid (153 mg, 93%): ¹H NMR (DMSO-d₆) δ 2.81 (br s, 4H), 3.28 (m,8H), 3.61 (br s, 4H), 3.85 (s, 2H), 4.27 (s, 2H), 4.36 (s, 2H), 7.29 (d,2H, J=7.7 Hz), 7.36 (d, 2H, J=7.7 Hz), 7.53 (m, 3H), 7.63 (d, 1H, J=7.7Hz), 7.80 (t, 1H, J=7.7 Hz), 7.99 (m, 1H), 8.69 (d, 1H, J=5.3 Hz);FAB-MS m/z 527 (MH+H⁸¹Br), 525 (MH+H⁷⁹Br), 445 (M+H). Anal. Calcd forC₂₇H₃₆N₆.6HBr.3H₂O: C, 32.95; H, 4.92; N, 8.54; Br, 48.72. Found: C,32.75; H, 4.89; N, 8.39; Br, 48.61.

EXAMPLE 15N-[7-(4,7,10-Triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD7051)

[0133] 1,3-Phenylenebis(ethylene)diamine

[0134] To a solution of 1,3-phenylenediacetonitrile (9.37 g, 60 mmol) inCH₃OH (saturated with NH₃, 150 mL) was added Raney-Ni (ca. 20 g,previously washed with CH₃OH several times) and the mixture washydrogenated at 45 psi on a Parr apparatus for 48 h. The reactionmixture was filtered through celite and the filtrate evaporated to givethe crude product (9.45 g, 96%) as a light green oil: ¹H NMR (CDCl₃) δ0.80-1.50 (br s, 4H), 2.70-2.76 (m, 4H), 2.94-2.99 (m, 4H), 7.01-7.07(m, 3H), 7.18-7.26 (m, 1H). This was used in the next step withoutfurther purification.

[0135]N,N′-Bis(2-Nitrobenzenesulfonyl)-1,3-Phenylenebis(ethylene)diamine

[0136] A solution of 2-nitrobenzenesulfonylchloride (19.94 g, 0.090 mol)in dry CH₂Cl₂ (70 mL) was added dropwise via cannula to a stirredsolution of 1,3-phenylenebis(ethylene)diamine (4.92 g, 0.030 mol) andEt₃N (16.7 mL, 0.12 mol) in dry CH₂Cl₂ (80 mL) under nitrogen. Thereaction mixture was stirred overnight at room temperature, and thenquenched with water (20 mL). The precipitate was collected by filtrationand washed with H₂O, CH₃OH, and Et₂O to give the desired product (9.22g, 58%) as a white solid: ¹H NMR (DMSO-d₆) δ 2.66 (t, 4H, J=7.7 Hz),3.08-3.18 (br s, 4H), 6.94 (d, 2H, J=6.4 Hz), 6.98 (s, 1H), 7.12 (dd,1H, J=6.4, 6.4 Hz), 7.78-7.84 (br m, 4H), 7.90-7.64 (br m, 4H), 8.16 (brs, 2H).

[0137]7-Diethoxyphosphoryl-4,10-bis(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-triene

[0138] Using General Procedure D: Reaction ofN,N′-bis(2-nitrobenzenesulfonyl)-1,3-phenylenebis(ethylene)diamine (8.74g, 16.4 mmol) withN-(diethoxyphosphoryl)-O,O′-bis(2-methylsulfonyl)di-ethanolamine (6.50g, 16.4 mmol) followed by silica gel column purification of the reactionproducts (1:15:35 CH₃OH-Et₂O-CH₂Cl₂) gave the title compound (4.03 g,33%) as a yellow foam: ¹H NMR (CDCl₃) δ 1.21 (t, 6H, J=6.4 Hz),2.39-2.46 (br m, 4H), 2.83-2.97 (br m, 8H), 3.68-3.72 (m, 4H), 3.80-3.92(m, 4H), 7.16 (d, 2H, J=6.5 Hz), 7.18 (s, 1H), 7.24 (dd, 1H, J=6.5, 6.5Hz), 7.60-7.74 (m, 6H), 8.04-8.08 (m, 2H).

[0139]4,10-Bis(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-triene

[0140] Using General Procedure E: Reaction of7-diethoxyphosphoryl-4,10-bis(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-triene(1.27 g, 1.72 mmol) followed by silica gel column purification of thereaction products (1:15:25 CH₃OH-EtOAc-CH₂Cl₂ then 20% CH₃OH in CH₂Cl₂)gave the title compound (574 mg, 57%) as a light yellow foam: ¹H NMR(CDCl₃) δ 1.42-1.50 (br, 1H), 2.01 (t, 4H, J=5.4 Hz), 2.90-3.10 (br m,4H), 3.08 (t, 4H, J=5.4 Hz), 3.56-3.60 (br m, 4H), 7.16 (d, 2H, J=6.8Hz), 7.31 (dd, 1H, J=6.8, 6.8 Hz), 7.36 (s, 1H), 7.61-7.63 (m, 2H),7.70-7.73 (m, 4H), 8.01-8.04 (m, 2H).

[0141]N-[7-[4,10-Bis(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-Nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0142] Using General Procedure F: Reaction of4,10-bis(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-triene(420 mg, 0.7 mmol) withN-[1-methylene-4-(chloromethylene)phenylene]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(302 mg, 0.7 mmol) followed by silica gel column purification of thereaction products (1:3 Et₂O-CH₂Cl₂) gave the title compound (491 mg,70%) as a pale yellow solid: ¹H NMR (CDCl₃) δ 1.97-2.02 (br m, 4H),2.73-2.78 (br m, 4H), 2.90-2.94 (br m, 4H), 3.32 (s, 2H), 3.64-3.67 (brm, 4H), 4.55 (s, 2H), 4.58 (s, 2H) 6.93 (d, 2H, J=8.0 Hz), 7.04 (d, 2H,J=8.0 Hz), 7.09-7.16 (br m, 4H), 7.23 (s, 1H), 7.29 (dd, 1H, J=7.9, 7.9Hz), 7.51-7.72 (m, 10H), 7.80-7.83 (m, 2H), 7.98 (d, 1H, J=7.8 Hz), 8.39(m, 1H).

[0143]N-[7-(4,7,10-Triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridinePentahydrobromide Dihydrate

[0144] Using General Procedure G: Reaction ofN-[7-[4,10-bis(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(380 mg, 0.38 mmol) followed by basic alumina column purification of thereaction products (1:20 CH₃OH-CH₂Cl₂) gave the free base of the titlecompound.

[0145] Conversion of the free base to the hydrobromide salt using asaturated solution of HBr(g) in CH₃OH followed by drying in vacuoovernight, gave the title compound (110 mg, 34% overall) as a whitesolid: ¹H NMR (DMSO-d₆) δ 2.80-2.88 (br s, 4H), 3.02-3.06 (br s, 4H),3.10-3.16 (br s, 4H), 3.38-3.44 (br s, 4H), 3.80-3.86 (br s, 2H),4.25-4.30 (br s, 2H), 4.33-4.37 (br s, 2H), 7.27-7.32 (br m, 4H),7.42-7.63 (br m, 6H), 7.96 (dd, 1H, J=7.7, 7.7 Hz), 8.10-8.30 (br s,3H), 8.69 (d, 1H, J=4.9 Hz), 9.45-9.62 (br s, 2H); FAB-MS m/z 526(MH+H⁸¹Br), 524 (MH+H⁷⁹Br), 444 (M+H, 100); Anal. Calcd forC₂₈H₄₂N₅Br₅.2H₂O: C, 38.03; H, 5.24; N, 7.92; Br, 45.18. Found: C,38.37; H, 5.28; N, 7.76; Br, 45.36.

EXAMPLE 16N-[1-(1,4,7-Triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7059) General Procedure H4-Diethoxyphosphoryl-7-(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetradecane

[0146] To a stirred solution of4-diethoxyphosphoryl-1,7-bis(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetradecane(1.32 g, 1.87 mmol) and K₂CO₃ (654 mg, 4.73 mmol) in DMF (11 mL) undernitrogen was added dropwise a solution of thiophenol (0.15 mL, 1.46mmol) in DMF (8 mL) over 1 h. The mixture was stirred for an additional3 h and then concentrated in vacuo. The residue was partitioned betweenCHCl₃ (50 mL) and H₂O (25 mL). The aqueous phase was separated andextracted with CHCl₃ (3×20 mL) and the combined organic extracts weredried (MgSO₄) and concentrated in vacuo. The residue was purified bycolumn chromatography on basic alumina (CHCl₃ then 3% MeOH/CHCl₃) togive the title compound as a yellow oil (178 mg, 23%): ¹H NMR (CDCl₃) δ1.31 (t, 6H, J=7.0 Hz), 1.40-1.67 (m, 10H), 2.65 (m, 2H), 2.78 (m, 2H),3.12 (m, 2H), 3.26-3.37 (m, 4H), 3.48 (m, 2H), 3.97-4.09 (m, 4H), 7.61(m, 1H), 7.68 (m, 214), 8.06 (m, 1H).

[0147]N-[1-[4-Diethoxyphosphoryl-7-(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetra-decanyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0148] Using General Procedure F: Reaction of4-diethoxyphosphoryl-7-(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetradecane(236 mg, 0.453 mmol) andN-[1-methylene-4-(chloromethylene)phenylene]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(238 mg, 0.551 mmol) followed by silica gel column purification of thereaction products (50% then 80% THF/hexane) gave the title compound as ayellow amorphous solid (305 mg, 73%): ¹H NMR (CDCl₃) δ 1.27 (t, 6H,J=7.1 Hz), 1.43 (br s, 8H), 1.63 (br s, 2H), 2.32 (br s, 2H), 2.55 (m,2H), 3.13-3.41 (m, 8H), 3.49 (s, 2H), 3.85-4.02 (m, 4H), 4.57 (s, 2H),4.58 (s, 2H), 7.07-7.22 (m, 6H), 7.51-7.71 (m, 7H), 7.98 (d, 1H, J=7.4Hz), 8.04 (m, 1H), 8.41 (d, 1H, J=4.0 Hz).

[0149]N-[1-[7-(2-Nitrobenzenesulfonyl)-1,4,7-triazacyclotetra-decanyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0150] Using General Procedure E: Reaction ofN-[1-[4-diethoxyphosphoryl-7-(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetra-decanyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(300 mg, 0.328 mmol) gave the title compound as a yellow amorphous solid(214 mg, 84%): ¹H NMR (CDCl₃) δ 1.34-1.44 (m, 8H), 1.69 (br s, 2H), 2.34(m, 2H), 2.52 (m, 2H), 2.62 (m, 2H), 2.82 (m, 2H), 3.42 (m, 6H), 4.58(s, 2H), 4.59 (s, 2H), 7.08-7.24 (m, 6H), 7.52-7.71 (m, 7H), 8.01 (m,2H), 8.42 (d, 1H, J=4.1 Hz).

[0151]N-[1-(1,4,7-Triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridinePentahydrobromide Dihydrate

[0152] A mixture ofN-[1-[7-(2-nitrobenzenesulfonyl)-1,4,7-triazacyclotetra-decanyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(209 mg, 0.268 mmol), K₂CO₃ (298 mg, 2.16 mmol), and thiophenol (0.17mL, 1.7 mmol) in acetonitrile (3 mL) were heated to 50° C. for 16.5 h.The reaction mixture was diluted with CH₂Cl₂ (10 mL) and washed withbrine (10 mL). The separated aqueous phase was extracted with CH₂Cl₂(3×5 mL) and the combined organic extracts were dried (MgSO₄) andevaporated. The residue was purified by column chromatography on basicalumina (CHCl₃ then 10% MeOH/CHCl₃) to give the free base of titlecompound as a yellow oil (92 mg, 84%): ¹H NMR (CDCl₃) δ 1.21-1.62 (m,10H), 2.40-2.49 (m, 4H), 2.60 (m, 6H), 2.79 (m, 2H), 3.49 (s, 2H), 3.80(s, 2H), 3.91 (s, 2H), 7.14 (m, 1H), 7.28 (m, 5H), 7.62 (m, 1H), 8.54(d, 1H, J=4.4 Hz).

[0153] Conversion of the free base (86 mg, 0.21 mmol) to thehydrobromide salt using a saturated solution of HBr(g) in MeOH (SeeGeneral Procedure G) followed by drying in vacuo at 40° C. for 15.5 hgave the title compound as a white solid (128 mg, 70%): ¹H NMR (D₂O) δ1.48 (br s, 6H), 1.82 (m, 4H), 3.22-3.36 (m, 10H), 3.50 (br s, 2H), 4.48(s, 4H), 4.64 (s, 2H), 7.62 (s, 4H), 7.88 (m, 1H), 7.94 (d, 1H, J=8.0Hz), 8.38 (m, 1H), 8.77 (d, 1H, J=5.2 Hz); FAB-MS m/z 492 (MH+H⁸¹Br),490 (M+H⁷⁹Br), 410 (M+H). Anal. Calcd for C₂₅H₃₉N₅.5HBr.2.5H₂O.0.1Et₂O:C, 35.20; H, 5.82; N, 8.08; Br, 46.10. Found: C, 35.48; H, 5.66; N,8.10; Br, 46.05.

EXAMPLE 17N-[4-[4,7,10,17-Tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7063)

[0154]7-Diethoxyphosphoryl-10-(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene

[0155] Using General Procedure H: Reaction of7-diethoxyphosphoryl-4,10-bis(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene(1.48 g, 2.00 mmol) with thiophenol (with an additional heating of thereaction mixture to 50° C. for 1.5 h after the addition) followed bysilica gel column purification of the reaction products (8% MeOH/CHCl₃)gave the title compound as a light yellow oil (423 mg, 52%): ¹H NMR(CDCl₃) δ 1.23 (t, 6H, J=7.0 Hz), 2.50 (br s, 2H), 2.79 (br s, 2H),3.02-3.15 (m, 10H), 3.82-3.98 (m, 6H), 7.06 (d, 2H, J=7.6 Hz), 7.54-7.63(m, 2H), 7.70 (m, 2H), 8.01 (br s, 1H).

[0156]N-[4-[7-Diethoxyphosphoryl-10-(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0157] Using General Procedure F: Reaction of7-diethoxyphosphoryl-10-(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene(410 mg, 0.738 mmol) andN-[1-methylene-4-(chloromethylene)phenylene]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(397 mg, 0.919 mmol) followed by silica gel column purification of thereaction products (50%, 80%, and 90% THF/hexane) gave the title compoundas a white amorphous solid (441 mg, 63%): ¹H NMR (CDCl₃) δ 1.15 (t, 6H,J=7.0 Hz), 2.42 (m, 4H), 2.77 (m, 2H), 2.92-3.02 (m, 6H), 3.10 (m, 2H),3.59 (s, 2H), 3.66-3.91 (m, 6H), 4.58 (s, 2H), 4.59 (s, 2H), 6.94 (d,1H, J=7.6 Hz), 7.07-7.14 (m, 6H), 7.22 (d, 1H, J=7.8 Hz), 7.51-7.72 (m,8H), 8.00 (d, 1H, J=7.8 Hz), 8.04 (m, 1H), 8.42 (d, 1H, J=4.0 Hz).

[0158]N-[4-[7-Diethoxyphosphoryl-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-2-(aminomethyl)pyridine

[0159] A mixture ofN-[4-[7-diethoxyphosphoryl-10-(2-nitrobenzenesulfonyl)-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(434 mg, 0.456 mmol), K₂CO₃ (508 mg, 3.68 mmol), and thiophenol (0.28mL, 2.7 mmol) were heated to 50° C. in CH₃CN (3.5 mL) under nitrogen for15 h. Upon cooling, the reaction mixture was partitioned between CHCl₃(15 mL) and brine (15 mL) and the aqueous layer was separated andextracted with CHCl₃ (3×5 mL). The combined organic extracts were dried(MgSO₄) and concentrated in vacuo and the residue was purified by columnchromatography on basic alumina (CHCl₃ then 10% MeOH/CHCl₃) to give ayellow oil (218 mg, 82%): ¹H NMR (CDCl₃) δ 1.16 (t, 6H, J=7.1 Hz), 2.35(m, 2H), 2.55 (m, 2H), 2.75 (m, 2H), 2.82 (m, 2H), 2.96-3.08 (m, 6H),3.16 (m, 2H), 3.68 (s, 2H), 3.69-3.88 (m, 4H), 3.82 (s, 2H), 3.93 (s,2H), 6.95 (d, 1H, J=7.6 Hz), 7.00 (d, 1H, J=7.5 Hz), 7.15-7.34 (m, 6H),7.50 (m, 1H), 7.65 (m, 1H), 8.56 (d, 1H, J=4.7 Hz).

[0160]N-[4-[4,7,10,17-Tetraazabicyclo[13.3.1]heptadeca-1(17)13,15-trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridineHexahydrobromide Hydrate

[0161] To a stirred solution ofN-[4-[7-diethoxyphosphoryl-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-2-(aminomethyl)pyridine(211 mg, 0.36 mmol) in acetic acid (0.6 mL) was added a freshly preparedsolution of saturated HBr(g) in acetic acid (6 mL) and the reactionmixture was allowed to stir at room temperature for 4 h. Addition ofdiethyl ether (10 mL) gave a white precipitate that was allowed tosettle to the bottom of the flask and the supernatant solution wasdecanted. The solid was washed by decantation with MeOH (4×5 mL) andether (6×5 mL) and the remaining traces of ether were removed byevaporation under reduced pressure. The product was dried in vacuo at40° C. for 17 h, to give the title compound as a pale yellow solid (223mg, 63%): ¹H NMR (D₂O) δ 3.14-3.36 (m, 10H), 3.55 (m, 4H), 3.75 (m, 2H),4.45 (s, 2H), 4.50 (s, 2H), 4.64 (s, 2H), 7.22 (m, 2H), 7.53 (s, 4H),7.70 (m, 1H), 7.95 (m, 1H), 8.00 (d, 1H, J=7.9 Hz), 8.46 (m, 1H), 8.79(d, 1H, J=3.9 Hz); FAB-MS m/z 527 (MH+H⁸¹Br), 525 (MH+H⁷⁹Br), 445 (M+H).Anal. Calcd for C₂₇H₃₆N₆.6HBr.1.5H₂O.0.2Et₂O: C, 34.35; H, 4.87; N,8.65; Br, 49.33. Found: C, 34.57; H, 5.04; N, 8.68; Br, 49.09.

EXAMPLE 18N-[4-[4,7,10-Triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD7058)

[0162]7-Diethoxyphosphoryl-10-(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-triene

[0163] Using General Procedure H: Reaction of7-diethoxyphosphoryl-4,10-bis(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-triene(1.11 g, 1.5 mmol) followed by silica gel column purification of thereaction products (2:5:20 CH₃OH-Et₂O-CH₂Cl₂ then 1:5 CH₃OH-CH₂Cl₂) gavethe title compound as a pale yellow oil (300 mg, 54%): ¹H NMR (CDCl₃) δ1.21 (t, 6H, J=7.1 Hz), 1.78-1.92 (br s, 1H), 2.31-2.38 (br m, 2H),2.56-2.60 (br m, 2H), 2.81-2.98 (br m, 10H), 3.60-3.64 (br m, 2H),3.75-3.91 (m, 4H), 7.05-7.12 (m, 2H), 7.24-7.29 (m, 2H), 7.60-7.63 (m,1H), 7.68-7.71 (m, 2H), 8.02-8.06 (m, 1H).

[0164]N-[4-[7-Diethoxyphosphoryl-10-(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0165] Using General Procedure F: Reactionof7-diethoxyphosphoryl-10-(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-triene(290 mg, 0.52 mmol) withN-[1-methylene-4-(chloromethylene)phenylene]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(271 mg, 0.63 mmol) followed by silica gel column purification of thereaction products (1:12:12 CH₃OH-Et₂O-CH₂Cl₂) gave the title compound(298 mg, 60%) as a pale yellow solid: ¹H NMR (CDCl₃) δ 1.17 (t, 6H,J=7.0 Hz), 2.29-2.45 (br m, 4H), 2.55-2.65 (br m, 2H), 2.71-2.75 (br s,4H), 2.85-2.91 (br m, 2H), 2.96-2.98 (br m, 2H), 3.57 (s, 2H), 3.67-3.84(br m, 6H), 4.57-4.61 (br s, 4H), 7.07-7.28 (br m, 10H), 7.55-7.71 (brm, 7H), 7.99-8.02 (m, 2H), 8.42-8.46 (m, 1H).

[0166]N-[4-[10-(2-Nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine

[0167] Using General Procedure E: Reaction ofN-[4-[7-diethoxyphosphoryl-10-(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(290 mg, 0.31 mmol) gave the title compound (240 mg, 95%) as a whitesolid: ¹H NMR (CDCl₃) δ 1.65-1.79 (br s, 1H, coincide with H₂O peak),2.15-2.19(br m, 4H), 2.44-2.48 (br m, 2H), 2.61-2.65 (br m, 2H),2.67-2.71 (br m, 2H), 3.00-3.04 (br m, 2H), 3.10-3.14 (br m, 2H),3.56-3.60 (br s, 4H), 4.55 (s, 2H), 4.61 (s, 2H), 6.96 (d, 1H, J=7.8Hz), 7.02-7.10 (br m, 6H), 7.22-7.28 (br m, 3H), 7.52-7.72 (br m, 7H),7.96-7.99 (m, 2H), 8.42-8.46 (m, 1H). This was used without furtherpurification.

[0168]N-[4-[4,7,10-Triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridinePentahydrobromide Dihydrate

[0169] Using General Procedure G: Reaction ofN-[4-[10-(2-nitrobenzenesulfonyl)-4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-N-(2-nitrobenzenesulfonyl)-2-(aminomethyl)pyridine(236 mg, 0.29 mmol) followed by alumina column purification of thereaction products (1:99 CH₃OH-CH₂Cl₂ then 1:10 CH₃OH-CH₂Cl₂) gave thefree base of the title compound (111 mg, 86%) as a pale yellow oil: ¹HNMR (CDCl₃) δ 2.24-2.28 (br s, 3H), 2.43-2.50 (br m, 4H), 2.58-2.62 (brm, 2H), 2.73-2.79 (br m, 8H), 2.95-2.98 (br m, 2H), 3.50 (s, 2H), 3.77(s, 2H), 3.90 (s, 2H), 6.83-6.87 (br m, 3H), 7.05-7.33 (br m, 7H),7.63-7.67 (m, 1H), 8.54-8.56 (m, 1H).

[0170] Conversion of the free base (104 mg, 0.23 mmol) to thehydrobromide salt using a saturated solution of HBr(g) in CH₃OH followedby drying of the product in vacuo, gave the title compound (101 mg, 52%)as a white solid: ¹H NMR (D₂O) δ 2.90-2.94 (br m, 2H), 2.97-3.01 (br m,2H), 3.12-3.16 (br m, 2H), 3.17-3.21 (br m, 2H), 3.24-3.28 (br m, 4H),3.47-3.51 (br m, 2H), 3.57-3.61 (br m, 2H), 4.38-4.42 (m, 6H), 7.34-7.40(m, 2H), 7.46-7.60 (m, 8H), 7.90-7.94 (m, 1H), 8.58-8.62 (m, 1H); FAB-MSm/z 526 (MH+H⁸¹Br), 524 (MH+H⁷⁹Br), 444 (M+H, 100); Anal. Calcd. forC₂₈H₄₂N₅Br₅2.5H₂O: C, 37.65; H, 5.30; N, 7.84; Br, 44.73. Found: C,37.53; H, 5.26; N, 7.79; Br, 44.75. TABLE 2 Inhibition of mAb 12G5binding Compound IC₅₀ ^(a) (ng/ml) AMD3100 27 AMD3465 3 AMD7049 52AMD7050 1 AMD7051 7 AMD7058 >1000 AMD7059 >1000 AMD7063 9 SDF-1α^(b) 270

[0171] Each of the following compounds, including AMD 3484 (see FIG.1AB), were synthesized according to procedures in Bridger, et al., J.Med. Chem. (1995) 38:366-378; J. Med. Chem. (1996) 39:109-119 and U.S.Pat. No. 5,583,131, U.S. Pat. No. 5,698,546 and allowed copending U.S.application Ser. No. 08/659,500, which are each incorporated in theirentirety by reference herein. TABLE 3 % Inhibition of Ca²⁺ flux (conc)or Compound IC₅₀ ^(a) (ng/ml) AMD3100   5 AMD3465   1 AMD7049 100% (1μg/ml) AMD7050 100% (1 μg/ml) AMD7051 100% (1 μg/ml) AMD7058  44% (1μg/ml) AMD7059  36% (1 μg/ml) AMD7063 100% (1 μg/ml)

EXAMPLE 191-[2,6-Dimethoxypyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecaneTetrahydrobromide (AMD 7032)

[0172]¹H NMR (D₂O) δ 1.78 (m, 2H), 1.88-1.92 (m, 2H), 2.59-3.03 (m,16H), 3.60 (s, 2H), 3.91 (s, 6H), 6.44 (s, 2H); ¹³C NMR (D₂O) δ 26.75,27.91, 48.34, 49.21, 49.89, 50.96, 52.01, 52.86, 54.88, 57.15, 57.53,59.42, 142.65, 157.42, 166.42; FAB MS m/z 352 (M+H); Anal.(C₁₈H₃₃N₅O₂4.1HBr0.25H₂O); Calc. C, 31.44; H, 5.51; N, 10.18; Br, 47.64.Found C, 31.17; H, 5.61; N, 9.92; Br, 47.54.

EXAMPLE 201-[2-Chloropyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecaneTetrahydrochloride Monohydrate (AMD 7048)

[0173]¹H NMR (D₂O) δ 1.92 (m, 2H), 2.12 (m, 2H), 2.77-2.80 (m, 4H),2.96-3.39 (m, 12H), 3.85 (s, 2H), 7.33 (d, 1H, J=5.4 Hz), 7.44 (s, 1H),8.40 (d, 1H, J=5.4 Hz); ¹³C NMR (D₂O) δ 24.75, 27.59, 47.40, 47.55,49.11, 49.23, 52.12, 52.40, 53.81, 54.42, 56.98, 126.97, 128.30, 151.90,152.34, 153.78; FAB MS m/z 326 (M+H); Anal.(C₁₆H₂₈N₅Cl.4.2HCl.0.5HOAc.1.1H₂O); Calc. C, 38.61; H, 6.94; N, 13.24;Cl, 34.86. Found C, 38.63; H, 6.94; N, 13.52; Cl, 34.61.

EXAMPLE 211-[2,6-Dimethylpyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecanePentahydrobromide Dihydrate (AMD 7060)

[0174]¹H NMR (D₂O) δ 1.77 (m, 2H), 1.93 (m, 2H), 2.48 (s, 6H), 2.61-3.00(m, 16H), 3.61 (s, 2H), 7.07 (s, 2H); ¹³C NMR (D₂O) δ 25.30, 26.22,27.49, 47.75, 48.65, 49.43, 50.41, 51.58, 52.19, 54.09, 56.63, 58.46;FAB MS m/z 320 (M+H); Anal. (C₁₈H₃₃N₅.5HBr.0.5HOAc.1.7H₂O); Calc. C,29.08; H, 5.57; N, 8.92; Br, 50.91. Found C, 29.04; H, 5.60; N, 8.73;Br, 50.87.

EXAMPLE 221-[2-Methylpyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecanePentahydrobromide Dihydrate (AMD 7061)

[0175]¹H NMR (D₂O) δ 2.01-2.08 (m, 2H), 2.22 (m, 2H), 2.70-2.72 (m, 2H),2.77 (s, 3H), 2.91-2.92 (m, 2H), 3.33-3.52 (m, 12H), 4.00 (s, 2H),7.86-7.89 (m, 2H), 8.56 (d, 1H, J=5.7 Hz); FAB MS m/z 306 (M+H); Anal.(C₁₇H₃₁N₅.4.9HBr.0.3HOAc.2.1H₂O); Calc. C, 27.9; H, 5.49; N, 9.24; Br,51.67. Found C, 28.08; H, 5.50; N, 9.56; Br, 51.56.

EXAMPLE 231-[2,6-Dichloropyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecaneTrihydrochloride Bishydrate (AMD 3451)

[0176]¹H NMR (D₂O) δ 1.83-1.88 (m, 2H), 2.04-2.08 (m, 2H), 2.58-2.62 (m,2H), 2.79-2.81 (m, 2H), 3.12-3.44 (m, 12H), 3.69 (s, 2H), 7.30 (s, 2H);¹³C NMR (D₂O) □ 136.26, 37.69, 55.26, 56.18, 58.33, 58.56, 58.92, 59.23,63.57, 65.44, 70.72, 140.37, 166.14, 167.37; FAB MS m/z 360 (M+H). Anal.(C₁₆H₃₄N₅Cl₅O₂): Calc. C, 38.00; H, 6.78; N, 13.85; Cl, 35.05. Found: C,38.33; H, 6.42; N, 13.88; Cl, 35.43.

EXAMPLE 241-[2-Chloropyrid-5-yl(methylene)]-1,4,8,11-tetraazacyclotetradecaneTetrahydrochloride Hemihydrate (AMD 3454)

[0177]¹H NMR (D₂O) δ 1.96-2.09 (br m, 4H), 3.02-3.17 (m, 4H), 3.19-3.28(br m, 8H), 3.40 (s, 4H), 4.10 (s, 2H), 7.40 (d, 1H, J=8.2 Hz), 7.80 (d,1H, J=8.2 Hz), 8.27 (s, 1H); ¹³C NMR (D₂O) δ 19.36, 19.47, 38.17, 38.64,39.06, 41.74, 41.88, 42.18, 45.66, 48.29, 54.62, 125.59, 126.69, 142.79,150.77, 151.75; FAB-MS m/z 326 (M+H). Anal. Calcd forC₁₆H₂₈N₅Cl.4HCl.0.5H₂O: C, 39.98; H, 6.92; N, 14.57; Cl, 36.87. Found:C, 40.36; H, 7.06; N, 14.56; Cl, 36.92.

[0178] General Procedures A, B and C were used to prepare the followingcompounds:

EXAMPLE 25N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-purinePentahydrobromide DiHydrate (AMD3472)

[0179]¹H NMR (D₂O) δ 1.88-2.05 (br m, 4H), 3.06-3.22 ( br m, 8H),3.27-3.44 (br m, 8H), 4.22 (s, 2H), 5.59 (s, 2H), 7.29 (s, 4H), 8.80 (s,1H), 9.11 (s, 1H), 9.28 (s, 1H); ¹³C NMR (D₂O) δ 18.39, 19.25, 37.24,37.55, 37.71, 41.13, 41.37, 41.71, 44.41, 47.73, 54.87, 129.45, 131.81,132.53, 136.67, 140.96, 147.88, 152.46, 154.37; FAB-MS m/z 423 (M+H).Anal. Calcd for C₂₃H₃₄N₈.5HBr.2H₂O.0.5CH₃CO₂H: C, 32.27; H, 5.07; N,12.54; Br, 44.73. Found: C, 32.66; H, 4.81;N, 12.41; Br, 44.58.

EXAMPLE 261-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-4-phenylpiperazinePentahydrobromide Hydrate (AMD3526)

[0180]¹H NMR (D₂O) δ 1.88-2.06 (br m, 4H), 3.11-3.53 (br m, 24H), 4.30(s, 2H), 4.32 (s, 2H), 6.89-6.97 (m, 3H), 7.19-7.24 (m, 2H), 7.49 (s,4H); ¹³C NMR (D₂O) δ 18.74, 19.37, 37.34, 41.47, 41.76, 42.03, 44.31,47.45, 48.26, 51.16, 58.48, 59.29, 118.18, 122.34, 129.99, 130.37,131.53, 131.85, 132.62, 148.47; FAB-MS m/z 547 (M+H⁸¹Br), 545 (M+H⁷⁹Br),465 (M+H). Anal. Calcd for C₂₈H₄₄N₆.5HBr.H₂O: C, 37.90; H, 5.79; N,9.47; Br, 45.03. Found: C, 37.72; H, 5.98; N, 9.38; Br, 46.78. TABLE 4Compound IC₅₀ ^(a) (μg/mL) AMD3451 8.9 AMD3472 45.4 AMD3454 32.3 AMD352682 AMD3100 >100

[0181] The active compounds may be administered in the form of apharmaceutical composition formulated according to well known principlesand incorporating the compound, preferably in unit dose form, incombination with a pharmaceutically acceptable diluent, carrier orexcipient. Such compositions may be in the form of solutions orsuspensions for injection, or irrigation or be in capsule, tablet,dragee, or other solid composition or as a solution or suspension fororal administration or formulated into pessaries or suppositories orsustained release forms of any of the above for implantation. Suitablediluents, carriers, excipients and other components are well known. Itmay be desirable also to formulate a composition for topicaladministration such as an ointment or cream. The compounds of theinvention may be used, in the form of a composition or alone.

[0182] The pharmaceutical compositions according to the invention may beformulated in unit dosages determined in accordance with conventionalpharmacological methods, suitably to provide active compounds in thedosage range in humans of from 0.1 to 100 mg/kg body weight per day, ina single dose or in a number of smaller doses. Preferred dosage rangesare 1 to 30 mg/kg body weight per day intravenous (iv) orintraperitoneal (ip). Other active compounds may be used in thecompositions or such active compounds or supplemental therapy may beincluded in a course of treatment. The pharmaceutical compositions areuseful for treatment of a patient comprising an effective therapeuticamount of the novel compound, where said compound effectively binds to achemokine receptor.

[0183] The present invention further contemplates the use of thesecompositions in the manufacture of a medicament for the treatment ofHIV-infected patients and/or the treatment of a disease by theregulation of endothelial cell function and/or the treatment of adisease relating to vascularization of the gastrointestinal tract;hematopoiesis; or cerebellar development.

[0184] In a method for treating a patient infected with HIV, thepharmaceutical composition is administered to said patient as atherapeutically effective amount of a pharmaceutical composition in apharmaceutically acceptable carrier. In a method of treating a patientwith a disease related to the regulation of endothelial cell function,the pharmaceutical composition is administered to said patient as atherapeutically effective amount of a pharmaceutical composition in apharmaceutically acceptable carrier. The present invention furthercontemplates methods of treating a patient with a disease relating tovascularization of the gastrointestinal tract; hematopoiesis; orcerebellar development, by administering to said patient atherapeutically effective amount of a pharmaceutical composition in apharmaceutically acceptable carrier.

[0185] The present invention further contemplates a method of treating apatient with a disease relating to basal leukocyte trafficking or theextravasation and tissue infiltration of leukocytes in response toinciting antigens, by administering to said patient a therapeuticallyeffective amount of a pharmaceutical composition in a pharmaceuticallyacceptable carrier. The present method also contemplates treating apatient, by administering to said patient a therapeutically effectiveamount of a pharmaceutical composition in a pharmaceutically acceptablecarrier, wherein said compound effectively binds to a chemokinereceptor.

1. A method to modulate a chemokine receptor in a subject which methodcomprises administering to a subject in need of such modulation aneffective amount of a compound of formula (I),V-CR¹R²—Ar—CR³R⁴—N(R⁵)—(CR⁶R⁷)_(x)—R⁸  (I)wherein V is a cyclicpolyamine moiety having a total of 9 to 24 members and from 3 to 6optionally substituted amine nitrogens spaced from each other by two ormore optionally substituted carbon atoms, and which may optionallycomprise a fused aromatic or heteroaromatic ring; R¹ to R⁷ areindependently hydrogen, or straight, branched or cyclic C₁₋₆ alkyl; R⁸is a heterocyclic group, a substituted aromatic group, or a mercaptangroup; Ar is an aromatic ring or heteroaromatic ring each said ringbeing optionally substituted; x is 1 or 2; or the acid addition saltsand metal complexes thereof, or a pharmaceutically acceptablecomposition thereof.
 2. The method of claim 1, wherein the chemokinereceptor is the CXCR4 or CCR5 receptor.
 3. The method of claim 2,wherein the receptor is the CXCR4 receptor.
 4. The method of claim 2,wherein the receptor is the CCR5 receptor.
 5. The method of claim 2,wherein the modulation effects amelioration of a condition selected fromthe group consisting of deficiencies in vascularization of thegastrointestinal tract, deficiency in hematopoiesis, deficiency incerebellar development; abnormal basal leukocyte trafficking; abnormalextravasation and tissue infiltration of leukocytes in response toinciting antigens; and abnormal calcium ion transport.
 6. The method ofclaim 5, wherein the modulation effects amelioration of deficiencies invascularization of the gastrointestinal tract.
 7. The method of claim 5,wherein the modulation effects amelioration of deficiency inhematopoiesis.
 8. The method of claim 5, wherein the modulation effectsamelioration of deficiency in cerebellar development.
 9. The method ofclaim 5, wherein the modulation effects amelioration of abnormal basalleukocyte trafficking.
 10. The method of claim 5, wherein the modulationeffects amelioration of abnormal extravasation and tissue infiltrationof leukocytes in response to inciting antigens.
 11. The method of claim5, wherein the modulation effects amelioration of abnormal calcium iontransport.
 12. The method of claim 1, wherein V is a 14 to 20 memberedfused or unfused ring system.
 13. The method of claim 12, wherein V is a4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl system ora 1,4,7-triazacyclotetra-decanyl system or a4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl system, or a1,4,8,11-tetraazacyclotetradecanyl system.
 14. The method of claim 1,wherein any substituents on Ar are selected from the group consisting ofalkyl, aryl, amino, alkoxy, hydroxy, halogen, carboxyl and carboxamido.15. The method of claim 1, wherein R⁸ is selected from the groupconsisting of pyridine, pyrimidine, pyrazine, imidazole, thiophene,thiophenyl, aminobenzyl, piperidinyl, piperazinyl and a mercaptan group.16. The method of claim 1, wherein each of R¹-R⁷ is hydrogen.
 17. Themethod of claim 1, wherein the compound of formula (I) is selected fromthe group consisting of:N-[4-(1,4,7-Triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7049);N-[7-(4,7,10,17-Tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7050);N-[7-(4,7,10-Triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7051);N-[4-[4,7,10-Triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7058);N-[1-(1,4,7-Triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7059);N-[4-[4,7,10,17-Tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 7063);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine(AMD 3465);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-N-methyl-2-(aminomethyl)pyridine(AMD 3538);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-4-)aminomethyl)pyridine(AMD 3500);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-3-(aminomethyl)pyridine(AMD 3499);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-(2-amino-methyl-5-methyl)pyrazine(AMD 3498);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(aminoethyl)pyridine(AMD 3497);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(aminomethyl)thiophene(AMD 3516);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-(aminoethyl)mercaptan(AMD 3530);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-2-amino-benzylamine(AMD 3517);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-4-amino-benzylamine(AMD 3544);N-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-4-(aminoethyl)imidazole(AMD 3543); orN-[1,4,8,11-Tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-benzylamine(AMD 3529) or the acid addition salts and metal complexes thereof.